Fig. 1.

Mutated spastins decorate different subset of MTs in HeLa cells. (A) HeLa cells expressing mutated GFP-tagged spastins were fixed and stained for β-tubulin. Magnifications of typical patterns are shown in the insets (red outline boxes). The green outlined inset in M1Δ-GFP shows typical empty ring-shaped structures observed with this construct. Scale bars: 20 µm. (B) HeLa cells that co-expressed M1CY-Flag and M85CY-GFP were fixed and stained for β-tubulin (not shown in the overlay). Scale bar: 10 µm. (C) Quantification of the overlap between GFP and Flag staining. Acquired images were processed as described in the Materials and Methods, and colocalization analysis between M85CY-GFP and M1CY-Flag (number of cells analyzed=11) was performed with ImageJ software. Colocalization coefficients between M1CY-GFP and M1CY-Flag are shown for comparison (number of cells analyzed=13). Data are shown as mean±s.e.m. Significance was determined by one-way ANOVA, Dunnett's post test. ****P<0.001. (D) HeLa cells transfected with the M85CY-GFP complementary DNA (cDNA) were subjected to live imaging at 37°C for 2 h. Images were acquired every 5 min. Scale bar: 10 µm.